1999 Fiscal Year Final Research Report Summary
Identification of Behavior of Protein in Various Environments and Its Application to Downstream Processing
| Project/Area Number |
09650883
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Kanai University |
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Principal Investigator |
YAGI Hideharu Kansai University, Faculty of Engineering, Professor, 工学部, 教授 (70109891)
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| Project Period (FY) |
1997 – 1999
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| Keywords | Protein / Refolding / Hydrophobic interaction / Aggregation / Crystallization / Biopolymer / Dynamic membrane |
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| Research Abstract |
The separation of the target material from the impurities requires the recognition of difference in the behavior or morphology in the solution in addition to the physical properties. The behavior and morphology affect the biological activity of the protein or its components. The aggregation of protein molecules results in failure of refolding of denatured protein but forms the dynamic membrane which is expected as a recyclable membrane for ultrafiltration. The aggregation of molecules in the solution is also the first step of crystallization.
This research project deals with the following themes from the standpoint of identification of behavior of protein in various environments and its application to downstream processings :
1. Application of hydrophobic interaction to the separation of protein during refolding.
The adsorption characteristics were different between the native and denatured lysozyme. Reactivated lysozyme by semibatch refolding was recovered by the column chromatography el
… More
uted gradiently.
2. Refolding of denatured enzymes in semibatch system
The denatured protein solution was supplied into the renaturation solution at a constant flow rate. The recoveries of activity in a semibatch operation were higher than those in batch operations.
3. Characteristics of formation of dynamic membrane on porous ceramic support.
The dynamic membrane of ovalbumin with self-rejection over 0.9 was formed on the porous ceramic support. The addition of salt or inhibitor of gelation inhibited the formation of membrane and lost the already formed membrane.
4. Effect of coexinsting amino acids or impurity on the crystallization of L-aspartic acid.
Coexisting amino acids were included fairly in the product L-aspartic crystals and detoriarated the crystal habit.
5. Separation by two-step filtration of biopolymer formed by Bacillus subtilis.
The biopolymer, γ-PGA, formed in Bacillus subtilis cultivation was separated from the microorganism by microfiltration and recovered by subsequent ultrafiltration. The fouling for the microfiltration was improved by back flushing with the permeate. Less
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